Laboratory of Advanced Chemical Biology, Graduate School of Life Science, and Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, N21 W11, Kita-ku, Sapporo 001-0021, Japan.
Mol Cell Proteomics. 2010 Mar;9(3):523-37. doi: 10.1074/mcp.M900559-MCP200. Epub 2009 Dec 14.
Although various glycoforms appear to participate independently in multiple molecular interactions in cellular adhesion that contribute to embryogenesis and organogenesis, a full portrait of the glycome diversity and the effect of the structural variations of cellular glycoforms on individual cell stages in proliferation and differentiation remain unclear. Here we describe a novel concept for the characterization of dynamic glycoform alteration during cell differentiation by means of "glycoblotting-based cellular glycomics," the only method allowing for rapid and quantitative glycan analysis. We demonstrated that processes of dynamic cellular differentiation of mouse embryonic carcinoma cells, P19CL6 and P19C6, and mouse embryonic stem cells into cardiomyocytes or neural cells can be monitored and characterized quantitatively by profiling entire N-glycan structures of total cell glycoproteins. Whole N-glycans enriched and identified by the glycoblotting method (67 glycans for P19CL6, 75 glycans for P19C6, and 72 glycans for embryonic stem cells) were profiled and bar-coded quantitatively with respect to the ratio of subgroups composed of characteristic glycoforms, namely glycotypes.
尽管各种糖型似乎独立参与细胞黏附中的多种分子相互作用，从而有助于胚胎发生和器官发生，但细胞糖型的聚糖多样性及其结构变异对增殖和分化中单个细胞阶段的影响的全貌仍不清楚。在这里，我们通过“基于糖印迹的细胞糖组学”描述了一种用于描述细胞分化过程中动态糖型改变的新概念，这是唯一允许快速和定量糖分析的方法。我们证明，通过对总细胞糖蛋白的完整 N-糖链结构进行分析，可以监测和定量分析小鼠胚胎癌细胞 P19CL6 和 P19C6 以及小鼠胚胎干细胞向心肌细胞或神经细胞的动态细胞分化过程。通过糖印迹法富集和鉴定的全 N-聚糖（P19CL6 为 67 种聚糖、P19C6 为 75 种聚糖、胚胎干细胞为 72 种聚糖）根据由特征性糖型（即糖型）组成的亚组的比例进行了分析和定量编码。
